Objectives: . This study aimed to investigate the regulatory functions of gamma-aminobutyric acid (GABA)ergic neural circuits from the inferior colliculus (IC) to the medial geniculate body (MGB) in salicylate-induced tinnitus. Methods: . Mice were treated with salicylate to induce tinnitus, and tinnitus-like behaviors were evaluated via gap prepulse inhibition of acoustic startle. Using combined viral tracing methodologies, we identified and mapped the pathways and connections from the IC to the MGB. Furthermore, we employed Gq-coupled human M3 designer receptors exclusively activated by designer drugs (DREADDs) and Gi-coupled human M4 DREADDs to achieve targeted excitation or suppression of GABAergic neurons in the IC and MGB. Following the administration of clozapine N-oxide, which binds to these receptors, we modulated these neural circuits to assess their impact on tinnitus severity in a mouse model. Results: . Our findings demonstrated that mice exposed to salicylate exhibited tinnitus-like behaviors. GABAergic neurons projecting retrogradely from the MGB to the IC were primarily concentrated in the external nucleus of the IC. After clozapine N-oxide administration, chemogenetic activation of IC-MGB GABAergic neurons aggravated salicylate-induced tinnitus. Additionally, activation of GABAergic neurons between the IC and MGB induced the perception of tinnitus even without salicylate. However, chemogenetic inhibition of the IC-MGB GABAergic circuit did not reverse salicylate-induced tinnitus. Conclusion . These findings suggest that activation of the IC-MGB GABAergic neural circuit may contribute to tinnitus generation through a mechanism distinct from that of salicylate-induced tinnitus. This study provides novel insights into the mechanisms underlying tinnitus.

This study assessed the role and mechanism of the recombinant Bacillus Calmette-Gue?rin vaccine(rBCG)with c-di-AMP adjuvant in regulating metabolism and immunity in epididymal white adipose(eWAT)in mice.Male C57BL/6 mice were intravenously immunized with BCG and rBCG,and their body weights were monitored.eWAT was isolated from the mice,and the stromal vascular fractions(SVFs)cell number was counted with a hemocytometer.Sections of mouse adipose tissue were prepared,and the size,number,and morphology of eWAT adipocytes and crown-like structure(CLS)formation were compared under a microscope after HE staining.The transcription levels of lipid metabolism-associated factors,cytokines and aging-associated genes in each group were determined with qRT-PCR.The body weights of mice gradually increased after immunization with BCG and rBCG.The proportions of eWAT increased,and the SVFs cell number decreased,in rBCG immunized mice.HE staining indicated that BCG immunization promoted hyperplasia,whereas rBCG immunization promoted hypertrophy of eWAT adipocytes;moreover,both BCG and rBCG immunization induced CLS formation in eWAT.The qRT-PCR results indicated that rBCG immunization inhibited the expression of genes associated with lipolysis and energy expenditure in eWAT.BCG immunization had little effect on cytokine transcription,whereas rBCG significantly induced the transcription of IFN-γ and IL-1Ra,and inhibited that of IL-15 and IL-2,but did not induce the expression of aging-associated genes.Thus,rBCG immunization induced eWAT adipocyte hypertrophy,which was associated with the inhibition of eWAT lipolysis and the regulation of cytokine expression.

Objective To investigate the efficacy of Bifidobacterium triple live bacteria enteric-coated capsules in the adjuvant treatment of patientys with gestational diabetes mellitus(GDM)complicated with depression,and its effect on intestinal flora and inflammatory indicators.Methods A total of 168 patients with GDM complicated with depression were selected and randomly divided into the control group(81 cases,treated with psychological counseling,dietary control and appropriate exercise)and the observation group(87 cases,treated with Bifidobacterium trilactis enteric-coated capsules on the basis of the control group,2 capsules each time,twice a day).Both groups were treated for 1 month continuously.The therapeutic effects were evaluated after treatment.The depression degree of patients was evaluated by Self-Rating Depression Scale(SDS)before treatment and one month after treatment.The fasting blood glucose(FBG)and 2-hour postprandial blood glucose(2 hPG)of patients were detected.The intestinal flora was detected by fluorescence quantitative PCR.The inflammatory indicators serum tumor necrosis factor-α(TNF-α)and serum C-reactive protein(CRP)were detected by enzyme-linked immunosorbent assay and immunoturbidimetry.The neutrophil and lymphocyte counts in peripheral venous blood were detected by automatic blood analyzer,and the neutrophil-to-lymphocyte ratio(NLR)was calculated.The adverse drug reactions were recorded.Results The total effective rate was 96.55%in the observation group,which was higher than that of the control group(87.65%,P＜0.05).After 1 month of treatment,the SDS score,FBG,2 hPG,serum TNF-α,CRP and NLR were decreased in both groups,and those of the observation group were lower than the control group(P＜0.05).One month after treatment,there was no significant difference in the number of intestinal flora compared with that before treatment in the control group.The number of Lactobacillus and Bifidobacterium in the observation group was increased compared with that before treatment,while the number of Escherichia coli and Bacteroides fragilis decreased.After 1 month of treatment,the number of Lactobacillus and Bifidobacterium in the observation group was higher than that in the control group,while the number of Escherichia coli and Bacteroides fragilis was lower than that in the control group(P＜0.01).The adverse reaction rate of the observation group was 3.45%,and there was no significant difference compared with 1.23%of the control group.Conclusion Bifidobacterium triple live bacteria enteric-coated capsules has a good clinical effect in the adjuvant treatment of GDM with depression,which can effectively control blood glucose level,improve intestinal flora,reduce inflammatory reaction and has good safety.

This study was aimed at investigating the effects and mechanisms of Toxoplasma gondii type Ⅰ(RH strain)ROP16 protein on proliferation and the cell cycle in mouse alveolar macrophage MH-S cells.We constructed a Toxoplasma gondii type Ⅰ(RH)ROP16 overexpression lentivirus,transduced MH-S cells,and then screened cells with puromycin to obtain a cell line stably overexpressing ROP16Ⅰ.RT-qPCR and western blotting were used to verify expression effects,CCK-8 assays were used to detect cell proliferation activity,and flow cytometry was used to detect cell cycle changes.Western blotting and RT-qPCR were used to detect the expression levels of p53,p21,CDK6,Cyclin D1,STAT3,p-STAT3(Y705),and JAK1 proteins or genes,and immunofluorescence was used to detect the expression levels of ROP16Ⅰ and p-STAT3(Y705)and their subcellular co-localization in MH-S cells.ROP16Ⅰ protein and gene expression were detected in MH-S cells transduced with lentivirus for ROP16Ⅰ overexpression.CCK-8 assays revealed that ROP16Ⅰ promoted the proliferation of MH-S cells(P＜0.01)and enhanced cell viability.Flow cytometry revealed that ROP16Ⅰ overexpression decreased the G0/G1 phase and elevated the G2 and S phases of the cell cycle in MH-S cells(P＜0.01 or P＜0.05).Compared with the MH-S cell group and MH-S-empty vector group,the MH-S-ROP16 cell group showed lower expression of p53 and p21 proteins;higher expression of CDK6,Cyclin D1,p-STAT3(Y705),and JAK1 proteins;lower expression of p53 and p21 mRNAs;and higher expression of CDK6 and Cyclin D1 mRNAs(all P＜0.01).Immunofluorescence revealed that ROP16Ⅰ co-localized with p-STAT3(Y705)in the nucleus and surrounding cytoplasm.Therefore,Toxoplasma gondii type Ⅰ(RH)ROP16Ⅰ protein activates the JAK-STAT3 pathway;shortens the G0/G1 phase and lengthens the G2/S phase of the cell cycle;and promotes cell proliferation.These findings provide a theoretical basis for revealing the mechanism of immune evasion of Toxoplasma gondii,and lay a foundation for research on the prevention and treatment of Toxoplasma gondii pneumonia.

Objective To evaluate the efficacy and safety of a novel intravascular lithotripsy(IVL)balloon—Vesscrack shockwave balloon—for vascular preparation before stent implantation in patients with severe coronary artery calcification(CAC).Methods This was a prospective,single-arm,multicenter study conducted in China from June 2022 to October 2022.Patients with severe CAC were treated with the Vesscrack shockwave balloon for lesion preparation,followed by drug-eluting stent(DES)implantation.Of these,33 patients underwent optical coherence tomography(OCT).The primary endpoint was procedural success,defined as successful stent implantation with residual stenosis≤30％and the absence of in-hospital major adverse events,including cardiac death,target vessel-related myocardial infarction,or target lesion revascularization.Results A total of 170 patients[mean age:(65.9±7.9)years,116 males]were enrolled.After treatment with IVL and DES,the minimum lumen diameter increased significantly compared to baseline[(2.34±0.40)mm vs.(0.95±0.33)mm,P＜0.001],the degree of stenosis was significantly reduced[(13.24±6.60)％vs.(65.18±10.59)％,P＜0.001].Procedural success was achieved in 100％of cases,and device success was 98.8％.The 30-day patient-related cardiovascular clinical composite endpoint(POCE)rate was 0.0,with no target lesion failure,no confirmed or potential thrombotic events were observed.The shockwave energy generator demonstrated excellent stability and ease of use.Among the 33 patients assessed with OCT,after IVL intervention,the maximum calcified area of the lumen[(3.51±1.51)mm2 vs.(2.85±1.80)mm2,P＜0.001],and the minimum lumen area within the target lesion[(3.08±1.04)mm2 vs.(2.02±0.75)mm2,P＜0.001],and after DES intervention,the luminal area of the largest calcified site[(6.59±1.64)mm2 vs.(2.85±1.80)mm2,P＜0.001]and the minimum luminal area within the target lesion[(6.19±1.45)mm2 vs.(2.02±0.75)mm2,P＜0.001]were significantly increased,and the differences were statistically significant.Conclusions The Vesscrack shockwave balloon is effective and safe for vascular preparation in patients with severe CAC prior to stent implantation.It achieves significant calcified plaque modification,high procedural success rates,and minimal complications.

Objective To compare the pain relief and long-term clinical success rate of vital pulp therapy and root canal treatment in mature permanent teeth with carious irreversible pulpitis.Methods A total of 90 patients diagnosed with carious irreversible pulpitis in mature permanent teeth were collected at Shanghai Stomatological Hospital from Jan 2021 to Jun 2022.They were randomly divided into two groups:test group(n=45)undergoing vital pulp therapy(VPT)and control group(n=45)undergoing root canal treatment(RCT).Pain scores were recorded before treatment,24 hours after operation and 7 days after operation.We conducted clinical evaluation and imaging analysis at 1,6,and 12 months after the surgery,then compared the pain scores and treatment success rates between the two groups.Results Eighty-one patients,including 39 patients in group VPT aged(31.00±1.43)years old and 42 patients in group RCT aged(30.60±1.54)years old,received follow-up for more than 1 year,and the success rate of the test group and control was 97.44%and 95.24%.The pain degree of the two groups was reduced at 24 hours and 7 days after operation(P<0.05),and the pain score of the test group was reduced compared with that in the control group 7 days after operation(P<0.01).Conclusion Compared with root canal treatment,vital pulp therapy for mature permanent teeth with carious irreversible pulpitis can achieve good results in short-term pain evolution and long-term clinical success.

Objective:To investigate the pharmacological mechanism of Compound Xuanju Capsule in the treatment of erectile dysfunction(ED)by using network pharmacology and molecular docking technology.Methods:The active ingredients and targets of Compound Xuanju Capsule were screened using Traditional Chinese Medicine Systematic Pharmacology Database and Analysis Platform(TCMSP).TTD,OMIM,DrugBank and GeneCards databases were used to obtain genes related to ED,and the union of the results was taken as the disease genes of ED.The common target of drug and disease was taken as the potential target of Compound Xuanju Capsule in ED,and the drug-disease interaction network was constructed by using Cytoscape software.The protein-protein interaction(PPI)network was constructed by using String database,which was then imported into Cytoscape to identify the key target.Based on the drug-disease intersection genes,GO and KEGG enrichment analyses were performed to predict the relevant signaling pathways and molecular mechanisms of Compound Xuanju Capsule for the treatment of ED.Autodock software was used to perform molecular docking between the active ingredients and the core targets.Results:Forty chemical components of Compound Xuanju Capsule were screened,and 239 predicted targets were obtained.A total of 1 907 ED-related genes were screened,and 97 common targets were identified between Compound Xuanju Capsule and ED,among which the core targets included EGFR,ESR1,HIF1A,PTGS2,and STAT3.The signaling pathways obtained by KEGG enrichment analysis included calcium signaling pathway,HIF-1 signaling pathway,PI3K-Akt signaling pathway,cGMP-PKG signaling pathway,relaxin signaling pathway,Serotonergic synapse signaling pathway.The molecular docking results showed that there were molecular binding sites between the key active ingredients and the core targets with strong binding activity.Conclusion:Compound Xuanju Capsule may treat ED through multi-target pathways such as anti-inflamnmato-ry and improving cellular oxidative stress.

This study was aimed at investigating the effects and mechanisms of Toxoplasma gondii type Ⅰ(RH strain)ROP16 protein on proliferation and the cell cycle in mouse alveolar macrophage MH-S cells.We constructed a Toxoplasma gondii type Ⅰ(RH)ROP16 overexpression lentivirus,transduced MH-S cells,and then screened cells with puromycin to obtain a cell line stably overexpressing ROP16Ⅰ.RT-qPCR and western blotting were used to verify expression effects,CCK-8 assays were used to detect cell proliferation activity,and flow cytometry was used to detect cell cycle changes.Western blotting and RT-qPCR were used to detect the expression levels of p53,p21,CDK6,Cyclin D1,STAT3,p-STAT3(Y705),and JAK1 proteins or genes,and immunofluorescence was used to detect the expression levels of ROP16Ⅰ and p-STAT3(Y705)and their subcellular co-localization in MH-S cells.ROP16Ⅰ protein and gene expression were detected in MH-S cells transduced with lentivirus for ROP16Ⅰ overexpression.CCK-8 assays revealed that ROP16Ⅰ promoted the proliferation of MH-S cells(P＜0.01)and enhanced cell viability.Flow cytometry revealed that ROP16Ⅰ overexpression decreased the G0/G1 phase and elevated the G2 and S phases of the cell cycle in MH-S cells(P＜0.01 or P＜0.05).Compared with the MH-S cell group and MH-S-empty vector group,the MH-S-ROP16 cell group showed lower expression of p53 and p21 proteins;higher expression of CDK6,Cyclin D1,p-STAT3(Y705),and JAK1 proteins;lower expression of p53 and p21 mRNAs;and higher expression of CDK6 and Cyclin D1 mRNAs(all P＜0.01).Immunofluorescence revealed that ROP16Ⅰ co-localized with p-STAT3(Y705)in the nucleus and surrounding cytoplasm.Therefore,Toxoplasma gondii type Ⅰ(RH)ROP16Ⅰ protein activates the JAK-STAT3 pathway;shortens the G0/G1 phase and lengthens the G2/S phase of the cell cycle;and promotes cell proliferation.These findings provide a theoretical basis for revealing the mechanism of immune evasion of Toxoplasma gondii,and lay a foundation for research on the prevention and treatment of Toxoplasma gondii pneumonia.

Objective To evaluate the efficacy and safety of a novel intravascular lithotripsy(IVL)balloon—Vesscrack shockwave balloon—for vascular preparation before stent implantation in patients with severe coronary artery calcification(CAC).Methods This was a prospective,single-arm,multicenter study conducted in China from June 2022 to October 2022.Patients with severe CAC were treated with the Vesscrack shockwave balloon for lesion preparation,followed by drug-eluting stent(DES)implantation.Of these,33 patients underwent optical coherence tomography(OCT).The primary endpoint was procedural success,defined as successful stent implantation with residual stenosis≤30％and the absence of in-hospital major adverse events,including cardiac death,target vessel-related myocardial infarction,or target lesion revascularization.Results A total of 170 patients[mean age:(65.9±7.9)years,116 males]were enrolled.After treatment with IVL and DES,the minimum lumen diameter increased significantly compared to baseline[(2.34±0.40)mm vs.(0.95±0.33)mm,P＜0.001],the degree of stenosis was significantly reduced[(13.24±6.60)％vs.(65.18±10.59)％,P＜0.001].Procedural success was achieved in 100％of cases,and device success was 98.8％.The 30-day patient-related cardiovascular clinical composite endpoint(POCE)rate was 0.0,with no target lesion failure,no confirmed or potential thrombotic events were observed.The shockwave energy generator demonstrated excellent stability and ease of use.Among the 33 patients assessed with OCT,after IVL intervention,the maximum calcified area of the lumen[(3.51±1.51)mm2 vs.(2.85±1.80)mm2,P＜0.001],and the minimum lumen area within the target lesion[(3.08±1.04)mm2 vs.(2.02±0.75)mm2,P＜0.001],and after DES intervention,the luminal area of the largest calcified site[(6.59±1.64)mm2 vs.(2.85±1.80)mm2,P＜0.001]and the minimum luminal area within the target lesion[(6.19±1.45)mm2 vs.(2.02±0.75)mm2,P＜0.001]were significantly increased,and the differences were statistically significant.Conclusions The Vesscrack shockwave balloon is effective and safe for vascular preparation in patients with severe CAC prior to stent implantation.It achieves significant calcified plaque modification,high procedural success rates,and minimal complications.

Objective To compare the pain relief and long-term clinical success rate of vital pulp therapy and root canal treatment in mature permanent teeth with carious irreversible pulpitis.Methods A total of 90 patients diagnosed with carious irreversible pulpitis in mature permanent teeth were collected at Shanghai Stomatological Hospital from Jan 2021 to Jun 2022.They were randomly divided into two groups:test group(n=45)undergoing vital pulp therapy(VPT)and control group(n=45)undergoing root canal treatment(RCT).Pain scores were recorded before treatment,24 hours after operation and 7 days after operation.We conducted clinical evaluation and imaging analysis at 1,6,and 12 months after the surgery,then compared the pain scores and treatment success rates between the two groups.Results Eighty-one patients,including 39 patients in group VPT aged(31.00±1.43)years old and 42 patients in group RCT aged(30.60±1.54)years old,received follow-up for more than 1 year,and the success rate of the test group and control was 97.44%and 95.24%.The pain degree of the two groups was reduced at 24 hours and 7 days after operation(P<0.05),and the pain score of the test group was reduced compared with that in the control group 7 days after operation(P<0.01).Conclusion Compared with root canal treatment,vital pulp therapy for mature permanent teeth with carious irreversible pulpitis can achieve good results in short-term pain evolution and long-term clinical success.